TOXICITY STUDY OF PARE LEAF EXTRACTS (MOMORDICA CHARANTIA) TO CULEX PIPIENS MOSQUITO LARVAE
J. Islamic Pharm. 2017. 2(2). 13-24
13
TOXICITY STUDY OF PARE LEAF EXTRACTS (MOMORDICA CHARANTIA) TO
CULEX PIPIENS MOSQUITO LARVAE
1*Rahmi
Annisa, 2 Muhammad Amir,
2 Hadi
Kuncoro
1
Department of Pharmacy, Faculty of Medicine and Health Sciences
Universitas Islam Negeri Maulana Malik Ibrahim, Malang, Indonesia
2 Faculty of Pharmacy, Universitas Mulawarman, Samarinda, Indonesia
* Email : apt.rahmiannisa@gmail.com
ABSTRACT
This study aims to determine the yield, toxicity, and toxic concentration of crude extracts of
ethanol, n-hexane fraction extract, ethyl acetate fraction extract and n-butanol fractions extract pare
leaf (Momordica charantiaL.) larvae Culex pipiens. The method used to determine the toxicity of
extract larvasida test larvae Culex pipiens to obtain the LC99 values obtained using the analysis of
Reed and Muench. Extraction and fractionation processis known to yield a ethanol crude extract
obtained 8.86%, n-hexane fraction extract of 1%, ethyl acetate fraction extract 1.80% and n-butanol
fractions extract of 0.80% and the toxicity obtained test results that pare leaf (Momordica charantia L.)
have the potential toxicity larvae larvae of Culex pipiens as indicated by the value by Lethality
Concrentation 99 % (LC99) in the crude extract ethanol 7416.52 ppm, n-hexane fraction extract of
2564.48 ppm, ethyl acetate fraction extract 2702.09 ppm and n-butanol fractions extract of 2665.63
ppm.
Keywords: Pare leaf (Momordica charantia L.), toxicity, larvae Culex pipiens
INTRODUCTION
Since ancient times the people of Indonesia know and use medicinal plants as one of
the efforts to overcome health problems before formal health care with synthetic chemicals
touched the community. Utilization of pare leaves as one of the plants that can be used as
insecticide mosquito larvae larvae (larvasida) that can be obtained from natural materials.
Larvacide is an ingredient insecticides capable of inhibiting life cycle or killing the
larvae stage in their natural habitat or at potential habitat. A mosquito larvaside that effective
must have fast work persistent at various breeding places mosquitoes, either on clean water or
on polluted water (Haeni, 2008).
Mosquitoes are among the species of insects that receive great attention in human
health, because it has the potential as a vector in the transmission of a disease. Insect borne
diseases are a common problem in Indonesia. Efforts to control a lot of vector is done
J. Islamic Pharm., an open access journal
ISSN : 2527-6123
J. Islamic Pharm. 2017. 2(2). 13-24
14
spraying several types of insecticides. The use of chemicals that are less cautious can pose a
danger to human health that is shortness of breath when the smell is stinging and allergic to
the skin, causing environmental pollution problem, besides synthetic chemicals can cause
resisaten in mosquito (Araujo, et al., 2006). In order to achieve optimal effort and minimize
the impact on the environment, an appropriate method should be developed for different areas
of the environment. Therefore it is necessary to find another alternative that is more simple
and not detrimental to environmental health (Samsuhidayat and Hutapea, 1991).
MATERIAL AND METHOD
Materials
Pare leaf Extract (Momordica charantia L.), aquadest, etanol, n-heksana, etil asetat, n-butanol,
tween 80, larva nyamuk Culex pipiens instar III
Procedure
Sample Preparation leaf of Momordica charantia L were collected at Kutai Kartanegara
town, Indonesia. The leafs were. The leafs were cleaned and drined. The dried sample was
pulverized until 60 mesh.
Extractionof Maseration The air-dried sample (250 gr) was extracted using Ethanol 96%
(1000 mL) until 48 hours and filtrated. The residue obtained after extraction was reextracted
with Ethanol 96% (1000 mL) or until the filtrat was transparent. The extract were drained and
concentrated under reduce pressure using Rotary Evaporator (40-50oC). The ethanol extract
was allowed to dry and its contstant weight was recorded.
Fractionated extracts performed with liquid-solid fractionation method. In this fractionation
will be made -heksana n extract fraction, ethyl acetate and n -butanol.
Preparation of bio-indicators (Culex pipiens mosquito larvae)
Culex pipiens mosquito larvae selected by criteria such as size, shape, and color of the
uniform. In the third instar spines and funnel chest began to clear respiratory blackish
brown. The test larva used was instar larvae of instar mosquitoes III.
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J. Islamic Pharm. 2017. 2(2). 13-24
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Design of Larvicides Bioactivity Test of Pare Leaf Extract
Table 1. Treatment of variation concentration of pare leaf extract
Concentration variation Pare Leaf Extract (Momordica charantia L.)
Replication
1
2
1
KEK1U1
KEK2U1
KEK3U1
KEK4U1
KEK5U1
2
KEK1U2
KEK2U2
KEK3U2
KEK4U2
KEK5U2
3
KEK1U3
KEK2U3
KEK3U3
KEK4U3
KEK5U3
Information
KEK1U1
KH1U1
KEA1U1
KB1U1
3
4
5
: Treatment of the first crude extract concentration on the first replay
: Treatment concentration n-hexane fraction first in first repeat
: Treatment concentration of ethyl acetate fraction first in first repeat
: Treatment concentrations of n-butanol fraction first in first repeat
Larvicides bioactivity test of Pare Leaf Extract to Culex pipiens mosquito larvae
Entered 10 Culex pipiens mosquito larvae was added to the vial containing water up to 10
mL. In each vial was treated differently according to the prepared test group then covered
with aluminum foil and given a small hole above it. Observed, noting the number of Culex
pipiens mosquito larvae mortality after 24 hours. LC99 values are calculated crude extract
ethanol, n -heksana fraction, of ethyl acetate and n -butanol fraction pare leaf (Momordica
charantia L.) to Culex mosquito larvae mortality pipiens.
Analysis of Data The data was described with table and graph. The lethal dose of 99%
(LD99) was determined by Reed and Muench method.
RESULTS AND DISCUSSION
Extract and Fractionation of Pare Leaf Extract
Pare leaf extract (Momordica charantia L.) is a leaf pare that has beeb extracted by
maceration by maceration method using ethanol andwith bulbs of 250 g. The maceration
process causes the solvent to penetrate the cell wall and into the cell cavity containing the
active substance. The active substance will dissolve together with organic solvent in the
presence of different concentrations between the active substance solution inside the cell and
outside the cell, then the concentrated solution will be pushed out. The event will be repeated
J. Islamic Pharm., an open access journal
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J. Islamic Pharm. 2017. 2(2). 13-24
16
resulting in a concentration equilibrium between the solution outside and inside the cell. After
conducting the subsequent sampling, the concentration of the sample solution which is aimed
to obtain the concentrated extract. Concentration is done by using a rotary evaporator. This
method aims to compounds Metab secondary olit not damaged by high temperatures. To
avoid the use of temperature on the concentration used process vacuum pumps, resulting in
reduced pressure and the solvent will evaporate at a temperature below its boiling point
(Dervish, 2000).
Extracts were obtained from 250 g dry sample is 22:16 g, so that can know the
amount of the yield obtained is at 8.86. To identify the components of a mixture based on the
level of the polarity of a compound fractionation process is carried out. The purpose of this
fractionation is to see bandage dingan pare leafextract bioactivity larvicides against the
mosquito larvae Culex pipens. Rendemen analysis faction aims to know the percentage of
fraction produced from the extract by fractionation method. Results yield extracts and
fractions can be seen in Table 2 the following:
Table 2. The results yield leaf extract of pare leaf exract (Momordica charantia L.)
No
Extract
Extract
Rendemen I
Rendemen II
Rendemen III
Weight(g)
(%)
(%)
(%)
1
Ekstrak etanol
22.16
2.67
8.86
-
2
Fraksi n-heksana
2.50
0.30
1.00
13.16
3
Fraksi Etil asetat
4.50
0.54
1.80
23.68
4
Fraksi n-butanol
2.00
0.24
0.80
10.53
Larvicides bioactivity test of Pare Leaf Extract to Culex pipiens mosquito larvae
To
determine
the bioactivity
of
larvicides leaf extract of
pare
leaf
extract
(Momordica charantia L.) then tested the mosquito larvae mortality with the use of acute
toxicity tests. This test is designed to determine the toxic effect of a compound that will occur
in a short time or administration with a certain dose (Department of Therapeutic
Pharmacology, 2000). This test is a toxicology bioactivity test that aims to determine the
biological activity of the extract or fraction of plants by observing the death response in
experimental animals. The death of the experimental animal is considered as a response to the
influence of certain compounds. This test is important to obtain information regarding the
concentration relationship with death response of experimental animals (Colagate and Russel,
1993).
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J. Islamic Pharm. 2017. 2(2). 13-24
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Test the efficacy of the ethanol extract of pare leaf extract (Momordica charantia L.)
against larvae of Aedes aegypti has been investigated by Lianawati (2008). The results of
these studies conclude that the leaf extract of pare leaf extract (Momordica charantia L.) has
the ability to kill mosquito larvae of Aedes aegypti with LC 50 value of 654.24 ppm and the
LC 90 for 1588.40 ppm.. This study aimed at testing the rough ethanol extract, extract fraction
of n-hexane, ethyl acetate fraction extract and extract of n-butanol fraction of the Culex
pipiens mosquito larvae.
This
study
uses
the
mosquito Culex
pipiens larva the third instar. First instar
larvae are stable and resistant to changes in temperature and humidity (Hanti, 1996). The
mosquito larvae in the third instar is more perfect than the physiological and morphological
third instar larvae thus more resistant to the materials of larvicides. Therefore it can be
concluded that the concentration that kills 100% of larvae of mosquitoes instar III also can
kill mosquito larvae instar III and not vice versa.
Results of testing the bioactivity of larvicides ethanol crude extract obtained from the
leaves of bitter melon five concentration variations with 3 repetitions.The test results are
shown in Table 3 :
Table 3. The results of bioactivity test larvicides ka sar ethanol extract of the l pare leaf
extract (Momordica charantia L.) against Culex pipiens mosquito larvae
Concentration
Amount
Log
Accumulate
Concentration
3750 ppm
5000 ppm
6250 ppm
7500 ppm
8750 ppm
3.57
3.69
3.79
3.87
3.94
In Table 3 above shows
Die
Life
9
15
21
30
30
21
15
9
0
0
the
test
Ratio
to
Mortality
tally dead
(%)
Die
Life
Accumulate
Ratio x
(x)
(y)
x : (x+y)
100 %
9
24
45
75
105
45
24
9
0
0
0.1667
0.5000
0.8384
1
1
16.67
50.00
83.84
100
100
results
crude
extract
ethanol,
in
which
the lowest test concentration of 3750 ppm has been giving activity in killing mosquito
larvae Culex pipiens and the high concentration of 8750 ppm level Culex pipiens mosquito
larvae mortality give
effect
to
kill
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ISSN : 2527-6123
all
the
test
animals. This
is
in
accordance
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18
with Harborne (1987) which states that the higher the concentration is given, the higher the
death rate bioindikator.
The
influence
of
the
concentration
of
ethanol
crude
of
pare
leaf
extract (Momordica charantia L.) the death of Culex pipiens mosquito larvae in shows that
their effect on mortality of Culex pipiens concentration of crude extract ethanol. The results of
test analysis using the method of Reed and Muench LC 99 values obtained ethanol crude
extract of 7416.52 ppm. The increase of extract concentration is directly proportional to the
number of test animals that died and vice versa inversely to the number of live test
animals. Increasing
concentrations
melon (Momordicacharantia L.)
of
ethanol
provides
an
crude
extract
increased
of
ability
leaves
to
kill
of
bitter
mosquito
larvae Culex pipiens, this is because the more concentrated solution concentration, the more
substances contained in the leaf extract of pare leaf extract (Momordica charantia L.), which
means the more toxins consumed Culex pipiens mosquito larvae, so the number ofmosquito
larvae mortality is also higher. This is in accordance with Nurhayati (2005) states the more
concentrated solution concentration means more active ingredient which can interfere with
the metabolism of the test animals.
Larvicides bioactivity test of Pare Leaf fraction n-hexane to Culex pipiens mosquito
larvae
Results bioactivity test of Pare Leaf fraction n-hexane to Culex pipiens mosquito
larvae can be seen in Table 4 :
Table 4. Bioactivity test of Pare Leaf fraction n-hexane to Culex pipiens mosquito larvae
Concentration
Amount
Log
Concentration
Die
Life
Accumulate
Ratio
Mortality
totally dead
%
Die
Life
Accumulate
Ratio x
(x)
(y)
x: (x + y)
100%
750 ppm
2.87
18
12
18
29
0.2429
38.29
1250 ppm
2.99
21
9
39
17
0.6964
69.64
1750 ppm
3.24
25
5
64
8
0.8887
88.87
2250 ppm
3.35
27
3
91
3
0.9681
96.81
2750 ppm
3.44
30
0
121
0
1
100
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J. Islamic Pharm. 2017. 2(2). 13-24
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Effect of Pare Leaf fraction n-hexane the death of Culex pipiens in shows that increasing the
concentration of n-hexane fraction is directly proportional to the number of test animals that
die and instead is inversely proportional to the number of test animals were alive. Increasing
concentrations of n-hexane fraction leaves of Pare Leaf fraction n-hexane improve the ability
to kill mosquito larvae of Culex pipiens.
Analysis using the method of Reed and
Muench LC 99 values obtained n -heksana extract fraction of 2564.48 ppm.
Larvicides Bioactivity test of Pare Leaf Fraction Ethyl Acetate to Culex pipiens mosquito
Larvae
Results bioactivity test of Pare Leaf fraction ethyl acetate to Culex pipiens mosquito
larvae can be seen in Table 5 :
Table 5. Bioactivity test of Pare Leaf fraction Ethyl Acetat to Culex pipiens mosquito larvae
Amount
Concentration
Accumulate
Log
Concentration
Die
Life
Ratio
Mortality
totally dead
%
Die
Life
Accumulate
Ratio x
(x)
(y)
x: (x + y)
100%
750 ppm
2.87
6
24
6
72
0.0769
7.69
1250 ppm
2.99
9
21
15
48
0.2380
23.80
1750 ppm
3.24
15
15
30
27
0.5263
52.63
2250 ppm
3.35
18
12
48
12
0800
80.00
2750 ppm
3.44
30
0
78
0
1
100
Effect of Pare Leaf fraction ethyl acetate the death of Culex pipiens in shows that
increasing the concentration of ethyl acetate fraction is directly proportional to the number of
test animals that die and instead is inversely proportional to the number of test animals were
alive. Increasing concentrations of ethyl acetate fraction leaves of Pare Leaf fraction ethyl
acetate improve the ability to kill mosquito larvae of Culex pipiens. Analysis using the
method of Reed and Muench LC 99 values obtained ethyl acetate extract fraction of 2702.09
ppm.
Larvicides bioactivity test of Pare Leaf fraction n-butanol to Culex pipiens mosquito
larvae
Results bioactivity test of Pare Leaf fraction n-butanol to Culex pipiens mosquito
larvae can be seen in Table 6 :
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J. Islamic Pharm. 2017. 2(2). 13-24
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Table 6. Bioactivity test of Pare Leaf fraction n-butanol to Culex pipiens mosquito larvae
Concentration
Amount
Log
Concentration
Die
Life
Accumulate
Ratio totally
Mortality
dead
%
Die
Life
Accumulate
Ratio x
(x)
(y)
x: (x + y)
100%
750 ppm
2.87
9
21
9
60
0.0869
8.69
1250 ppm
2.99
12
18
21
39
0.3500
35.00
1750 ppm
3.24
15
15
36
21
0.6316
63.16
2250 ppm
3.35
24
6
60
6
0.9091
90.91
2750 ppm
3.44
30
0
90
0
1
100
Effect of Pare Leaf fraction n-butanol the death of Culex pipiens in shows that
increasing the concentration of n-butanol fraction is directly proportional to the number of test
animals that die and instead is inversely proportional to the number of test animals were
alive. Increasing concentrations of n-butanol fraction leaves of Pare Leaf fraction ethyl
acetate improve the ability to kill mosquito larvae of Culex pipiens. Analysis using the
method of Reed and Muench LC 99 values obtained ethyl acetate
extract fraction of
2665.63 ppm.
Based on these descriptions, it is known that the crude extract of ethanol and the
three extract fractions tested had as larvicidal activity, it is in accordance with Dalimartha
(2008), which explains that bitter melon plants can be used to kill insects. Pare leaf extract
and the fractions have the activity level is different, the difference in larvicidal activity of
extracts and fractions caused some fractionation process wherein the compound contained in
the extract chemical separation based on the degree of polarity so as to provide a
different larvicidal activity.
The content of compounds in bitter melon leaves are flavonoids, alkaloids, saponins
and terpenoids (Syamsuhidayat and Hutapea, 1991). Based on the study h acyl phytochemical
test extracts or extract fractions pare leaf by Hernawati (2006) in his research on the potential
p acre fruit (Momordica charantia L.) as a herbal ntifertilitas show that ethanol crude extract
contains alkaloids, saponins, flavonoids, and triterpenes oid. Fraction n-hexane containing
triterpenoids,flavonoids containing ethyl acetate and n -butanol containing flavonoids.
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J. Islamic Pharm. 2017. 2(2). 13-24
21
Fortunately (1993) and Kristanti (2008) explains that the insecticide can get into the
insect's body through various ways, among others: as a stomach poison(stomatch poison) that
goes into the insect's body through the digestive tract of insects, gastrointestinal tract
mosquitoes have a structure such as a wall body . Thus the absorption of insecticides in the
digestion equals absorption of the body wall. Contact poison (contact Poisoining) that enter
through the skin or body wall, dinding body part that can absorb large amounts of
insecticide. The body wall has a base layer that are semipermeable membrane so that it can
choose the types of compounds that can pass through and the last one fumigant or breathing
into the insect's body through the respiratory system. The respiratory tract is called the
trachea. Air and oxygen enter the trachea by diffusion assisted by abdominal
movement. Oxygen will be directly related to the network. Insecticides enter the respiratory
system in the form of gas or fine grains that are brought to the living tissue.
Function alkaloids, triterpenoids, saponins and flavonoids in the leaves of bitter
melon can hinder eating larvae (antifedant). These compounds can enter the digestive through
immersion extract concentration ingested by the larvae of the mosquito Culex pipiens, then
enter the digestive tract and are absorbed by the intestinal wall and then circulate along the
blood that would interfere with the metabolism of the body of the mosquito larvae Culex
pipiens that lack of energy for activities that lead to Mosquitoes finally die. The workings of
these compounds is to act as stomatch poisoning or stomach poison. Therefore, if these
compounds are included in the larval body, ala t be disturbed digestion (Cahyadi, 2009).
CONCLUSION
The crude extract of bitter melon leaves ethanol and fractions have an activity as larvicides
against the mosquito larvae Culex pipiens the LC 99 value of each is at 7416.52 ppm ethanol
extract, extract n -heksana fraction 2564.48 ppm, extract ethyl acetate fraction 2702.09 ppm
and extract n -butanol fraction 2665.63 ppm.
ACKNOWLEDGEMENTS
The development of this plant needs to be done because this plant is easy to find and
the number is many. E kstrak and pare leaf extract fraction having as larvicidal
activity. Extract fraction n -heksana known to have the most excellent larvicidal activity, these
results can be used as a reference in use are the most excellent naan extract as larvicides for
further research so that the development of further research on bitter melon leaves that leads
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to the use of these crops is necessary, the isolation of pure compound From leaf pare is a way
of development further to do in the future so that it can be used as a natural larvicidal derived
from leaf extracts of bitter melon (Momordica charantia L.).
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Masyarakat Universitas Diponegoro. Semarang
[33] Waren, K.S 1993. Immunology and Molecular Biology of Parasitic Infection 3rd.
Blackwell Scientific Publication. Boston
[34] Widiarti. 1991. Tinjauan Penelitian Pengendalian Vektor Malaria Secara Hayati dan
Pengelolaan Lingkungan. Pusat Penelitian Ekologi Badan LitBangKes DepKes RI.
Jakarta
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13
TOXICITY STUDY OF PARE LEAF EXTRACTS (MOMORDICA CHARANTIA) TO
CULEX PIPIENS MOSQUITO LARVAE
1*Rahmi
Annisa, 2 Muhammad Amir,
2 Hadi
Kuncoro
1
Department of Pharmacy, Faculty of Medicine and Health Sciences
Universitas Islam Negeri Maulana Malik Ibrahim, Malang, Indonesia
2 Faculty of Pharmacy, Universitas Mulawarman, Samarinda, Indonesia
* Email : apt.rahmiannisa@gmail.com
ABSTRACT
This study aims to determine the yield, toxicity, and toxic concentration of crude extracts of
ethanol, n-hexane fraction extract, ethyl acetate fraction extract and n-butanol fractions extract pare
leaf (Momordica charantiaL.) larvae Culex pipiens. The method used to determine the toxicity of
extract larvasida test larvae Culex pipiens to obtain the LC99 values obtained using the analysis of
Reed and Muench. Extraction and fractionation processis known to yield a ethanol crude extract
obtained 8.86%, n-hexane fraction extract of 1%, ethyl acetate fraction extract 1.80% and n-butanol
fractions extract of 0.80% and the toxicity obtained test results that pare leaf (Momordica charantia L.)
have the potential toxicity larvae larvae of Culex pipiens as indicated by the value by Lethality
Concrentation 99 % (LC99) in the crude extract ethanol 7416.52 ppm, n-hexane fraction extract of
2564.48 ppm, ethyl acetate fraction extract 2702.09 ppm and n-butanol fractions extract of 2665.63
ppm.
Keywords: Pare leaf (Momordica charantia L.), toxicity, larvae Culex pipiens
INTRODUCTION
Since ancient times the people of Indonesia know and use medicinal plants as one of
the efforts to overcome health problems before formal health care with synthetic chemicals
touched the community. Utilization of pare leaves as one of the plants that can be used as
insecticide mosquito larvae larvae (larvasida) that can be obtained from natural materials.
Larvacide is an ingredient insecticides capable of inhibiting life cycle or killing the
larvae stage in their natural habitat or at potential habitat. A mosquito larvaside that effective
must have fast work persistent at various breeding places mosquitoes, either on clean water or
on polluted water (Haeni, 2008).
Mosquitoes are among the species of insects that receive great attention in human
health, because it has the potential as a vector in the transmission of a disease. Insect borne
diseases are a common problem in Indonesia. Efforts to control a lot of vector is done
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J. Islamic Pharm. 2017. 2(2). 13-24
14
spraying several types of insecticides. The use of chemicals that are less cautious can pose a
danger to human health that is shortness of breath when the smell is stinging and allergic to
the skin, causing environmental pollution problem, besides synthetic chemicals can cause
resisaten in mosquito (Araujo, et al., 2006). In order to achieve optimal effort and minimize
the impact on the environment, an appropriate method should be developed for different areas
of the environment. Therefore it is necessary to find another alternative that is more simple
and not detrimental to environmental health (Samsuhidayat and Hutapea, 1991).
MATERIAL AND METHOD
Materials
Pare leaf Extract (Momordica charantia L.), aquadest, etanol, n-heksana, etil asetat, n-butanol,
tween 80, larva nyamuk Culex pipiens instar III
Procedure
Sample Preparation leaf of Momordica charantia L were collected at Kutai Kartanegara
town, Indonesia. The leafs were. The leafs were cleaned and drined. The dried sample was
pulverized until 60 mesh.
Extractionof Maseration The air-dried sample (250 gr) was extracted using Ethanol 96%
(1000 mL) until 48 hours and filtrated. The residue obtained after extraction was reextracted
with Ethanol 96% (1000 mL) or until the filtrat was transparent. The extract were drained and
concentrated under reduce pressure using Rotary Evaporator (40-50oC). The ethanol extract
was allowed to dry and its contstant weight was recorded.
Fractionated extracts performed with liquid-solid fractionation method. In this fractionation
will be made -heksana n extract fraction, ethyl acetate and n -butanol.
Preparation of bio-indicators (Culex pipiens mosquito larvae)
Culex pipiens mosquito larvae selected by criteria such as size, shape, and color of the
uniform. In the third instar spines and funnel chest began to clear respiratory blackish
brown. The test larva used was instar larvae of instar mosquitoes III.
J. Islamic Pharm., an open access journal
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J. Islamic Pharm. 2017. 2(2). 13-24
15
Design of Larvicides Bioactivity Test of Pare Leaf Extract
Table 1. Treatment of variation concentration of pare leaf extract
Concentration variation Pare Leaf Extract (Momordica charantia L.)
Replication
1
2
1
KEK1U1
KEK2U1
KEK3U1
KEK4U1
KEK5U1
2
KEK1U2
KEK2U2
KEK3U2
KEK4U2
KEK5U2
3
KEK1U3
KEK2U3
KEK3U3
KEK4U3
KEK5U3
Information
KEK1U1
KH1U1
KEA1U1
KB1U1
3
4
5
: Treatment of the first crude extract concentration on the first replay
: Treatment concentration n-hexane fraction first in first repeat
: Treatment concentration of ethyl acetate fraction first in first repeat
: Treatment concentrations of n-butanol fraction first in first repeat
Larvicides bioactivity test of Pare Leaf Extract to Culex pipiens mosquito larvae
Entered 10 Culex pipiens mosquito larvae was added to the vial containing water up to 10
mL. In each vial was treated differently according to the prepared test group then covered
with aluminum foil and given a small hole above it. Observed, noting the number of Culex
pipiens mosquito larvae mortality after 24 hours. LC99 values are calculated crude extract
ethanol, n -heksana fraction, of ethyl acetate and n -butanol fraction pare leaf (Momordica
charantia L.) to Culex mosquito larvae mortality pipiens.
Analysis of Data The data was described with table and graph. The lethal dose of 99%
(LD99) was determined by Reed and Muench method.
RESULTS AND DISCUSSION
Extract and Fractionation of Pare Leaf Extract
Pare leaf extract (Momordica charantia L.) is a leaf pare that has beeb extracted by
maceration by maceration method using ethanol andwith bulbs of 250 g. The maceration
process causes the solvent to penetrate the cell wall and into the cell cavity containing the
active substance. The active substance will dissolve together with organic solvent in the
presence of different concentrations between the active substance solution inside the cell and
outside the cell, then the concentrated solution will be pushed out. The event will be repeated
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J. Islamic Pharm. 2017. 2(2). 13-24
16
resulting in a concentration equilibrium between the solution outside and inside the cell. After
conducting the subsequent sampling, the concentration of the sample solution which is aimed
to obtain the concentrated extract. Concentration is done by using a rotary evaporator. This
method aims to compounds Metab secondary olit not damaged by high temperatures. To
avoid the use of temperature on the concentration used process vacuum pumps, resulting in
reduced pressure and the solvent will evaporate at a temperature below its boiling point
(Dervish, 2000).
Extracts were obtained from 250 g dry sample is 22:16 g, so that can know the
amount of the yield obtained is at 8.86. To identify the components of a mixture based on the
level of the polarity of a compound fractionation process is carried out. The purpose of this
fractionation is to see bandage dingan pare leafextract bioactivity larvicides against the
mosquito larvae Culex pipens. Rendemen analysis faction aims to know the percentage of
fraction produced from the extract by fractionation method. Results yield extracts and
fractions can be seen in Table 2 the following:
Table 2. The results yield leaf extract of pare leaf exract (Momordica charantia L.)
No
Extract
Extract
Rendemen I
Rendemen II
Rendemen III
Weight(g)
(%)
(%)
(%)
1
Ekstrak etanol
22.16
2.67
8.86
-
2
Fraksi n-heksana
2.50
0.30
1.00
13.16
3
Fraksi Etil asetat
4.50
0.54
1.80
23.68
4
Fraksi n-butanol
2.00
0.24
0.80
10.53
Larvicides bioactivity test of Pare Leaf Extract to Culex pipiens mosquito larvae
To
determine
the bioactivity
of
larvicides leaf extract of
pare
leaf
extract
(Momordica charantia L.) then tested the mosquito larvae mortality with the use of acute
toxicity tests. This test is designed to determine the toxic effect of a compound that will occur
in a short time or administration with a certain dose (Department of Therapeutic
Pharmacology, 2000). This test is a toxicology bioactivity test that aims to determine the
biological activity of the extract or fraction of plants by observing the death response in
experimental animals. The death of the experimental animal is considered as a response to the
influence of certain compounds. This test is important to obtain information regarding the
concentration relationship with death response of experimental animals (Colagate and Russel,
1993).
J. Islamic Pharm., an open access journal
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J. Islamic Pharm. 2017. 2(2). 13-24
17
Test the efficacy of the ethanol extract of pare leaf extract (Momordica charantia L.)
against larvae of Aedes aegypti has been investigated by Lianawati (2008). The results of
these studies conclude that the leaf extract of pare leaf extract (Momordica charantia L.) has
the ability to kill mosquito larvae of Aedes aegypti with LC 50 value of 654.24 ppm and the
LC 90 for 1588.40 ppm.. This study aimed at testing the rough ethanol extract, extract fraction
of n-hexane, ethyl acetate fraction extract and extract of n-butanol fraction of the Culex
pipiens mosquito larvae.
This
study
uses
the
mosquito Culex
pipiens larva the third instar. First instar
larvae are stable and resistant to changes in temperature and humidity (Hanti, 1996). The
mosquito larvae in the third instar is more perfect than the physiological and morphological
third instar larvae thus more resistant to the materials of larvicides. Therefore it can be
concluded that the concentration that kills 100% of larvae of mosquitoes instar III also can
kill mosquito larvae instar III and not vice versa.
Results of testing the bioactivity of larvicides ethanol crude extract obtained from the
leaves of bitter melon five concentration variations with 3 repetitions.The test results are
shown in Table 3 :
Table 3. The results of bioactivity test larvicides ka sar ethanol extract of the l pare leaf
extract (Momordica charantia L.) against Culex pipiens mosquito larvae
Concentration
Amount
Log
Accumulate
Concentration
3750 ppm
5000 ppm
6250 ppm
7500 ppm
8750 ppm
3.57
3.69
3.79
3.87
3.94
In Table 3 above shows
Die
Life
9
15
21
30
30
21
15
9
0
0
the
test
Ratio
to
Mortality
tally dead
(%)
Die
Life
Accumulate
Ratio x
(x)
(y)
x : (x+y)
100 %
9
24
45
75
105
45
24
9
0
0
0.1667
0.5000
0.8384
1
1
16.67
50.00
83.84
100
100
results
crude
extract
ethanol,
in
which
the lowest test concentration of 3750 ppm has been giving activity in killing mosquito
larvae Culex pipiens and the high concentration of 8750 ppm level Culex pipiens mosquito
larvae mortality give
effect
to
kill
J. Islamic Pharm., an open access journal
ISSN : 2527-6123
all
the
test
animals. This
is
in
accordance
J. Islamic Pharm. 2017. 2(2). 13-24
18
with Harborne (1987) which states that the higher the concentration is given, the higher the
death rate bioindikator.
The
influence
of
the
concentration
of
ethanol
crude
of
pare
leaf
extract (Momordica charantia L.) the death of Culex pipiens mosquito larvae in shows that
their effect on mortality of Culex pipiens concentration of crude extract ethanol. The results of
test analysis using the method of Reed and Muench LC 99 values obtained ethanol crude
extract of 7416.52 ppm. The increase of extract concentration is directly proportional to the
number of test animals that died and vice versa inversely to the number of live test
animals. Increasing
concentrations
melon (Momordicacharantia L.)
of
ethanol
provides
an
crude
extract
increased
of
ability
leaves
to
kill
of
bitter
mosquito
larvae Culex pipiens, this is because the more concentrated solution concentration, the more
substances contained in the leaf extract of pare leaf extract (Momordica charantia L.), which
means the more toxins consumed Culex pipiens mosquito larvae, so the number ofmosquito
larvae mortality is also higher. This is in accordance with Nurhayati (2005) states the more
concentrated solution concentration means more active ingredient which can interfere with
the metabolism of the test animals.
Larvicides bioactivity test of Pare Leaf fraction n-hexane to Culex pipiens mosquito
larvae
Results bioactivity test of Pare Leaf fraction n-hexane to Culex pipiens mosquito
larvae can be seen in Table 4 :
Table 4. Bioactivity test of Pare Leaf fraction n-hexane to Culex pipiens mosquito larvae
Concentration
Amount
Log
Concentration
Die
Life
Accumulate
Ratio
Mortality
totally dead
%
Die
Life
Accumulate
Ratio x
(x)
(y)
x: (x + y)
100%
750 ppm
2.87
18
12
18
29
0.2429
38.29
1250 ppm
2.99
21
9
39
17
0.6964
69.64
1750 ppm
3.24
25
5
64
8
0.8887
88.87
2250 ppm
3.35
27
3
91
3
0.9681
96.81
2750 ppm
3.44
30
0
121
0
1
100
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J. Islamic Pharm. 2017. 2(2). 13-24
19
Effect of Pare Leaf fraction n-hexane the death of Culex pipiens in shows that increasing the
concentration of n-hexane fraction is directly proportional to the number of test animals that
die and instead is inversely proportional to the number of test animals were alive. Increasing
concentrations of n-hexane fraction leaves of Pare Leaf fraction n-hexane improve the ability
to kill mosquito larvae of Culex pipiens.
Analysis using the method of Reed and
Muench LC 99 values obtained n -heksana extract fraction of 2564.48 ppm.
Larvicides Bioactivity test of Pare Leaf Fraction Ethyl Acetate to Culex pipiens mosquito
Larvae
Results bioactivity test of Pare Leaf fraction ethyl acetate to Culex pipiens mosquito
larvae can be seen in Table 5 :
Table 5. Bioactivity test of Pare Leaf fraction Ethyl Acetat to Culex pipiens mosquito larvae
Amount
Concentration
Accumulate
Log
Concentration
Die
Life
Ratio
Mortality
totally dead
%
Die
Life
Accumulate
Ratio x
(x)
(y)
x: (x + y)
100%
750 ppm
2.87
6
24
6
72
0.0769
7.69
1250 ppm
2.99
9
21
15
48
0.2380
23.80
1750 ppm
3.24
15
15
30
27
0.5263
52.63
2250 ppm
3.35
18
12
48
12
0800
80.00
2750 ppm
3.44
30
0
78
0
1
100
Effect of Pare Leaf fraction ethyl acetate the death of Culex pipiens in shows that
increasing the concentration of ethyl acetate fraction is directly proportional to the number of
test animals that die and instead is inversely proportional to the number of test animals were
alive. Increasing concentrations of ethyl acetate fraction leaves of Pare Leaf fraction ethyl
acetate improve the ability to kill mosquito larvae of Culex pipiens. Analysis using the
method of Reed and Muench LC 99 values obtained ethyl acetate extract fraction of 2702.09
ppm.
Larvicides bioactivity test of Pare Leaf fraction n-butanol to Culex pipiens mosquito
larvae
Results bioactivity test of Pare Leaf fraction n-butanol to Culex pipiens mosquito
larvae can be seen in Table 6 :
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J. Islamic Pharm. 2017. 2(2). 13-24
20
Table 6. Bioactivity test of Pare Leaf fraction n-butanol to Culex pipiens mosquito larvae
Concentration
Amount
Log
Concentration
Die
Life
Accumulate
Ratio totally
Mortality
dead
%
Die
Life
Accumulate
Ratio x
(x)
(y)
x: (x + y)
100%
750 ppm
2.87
9
21
9
60
0.0869
8.69
1250 ppm
2.99
12
18
21
39
0.3500
35.00
1750 ppm
3.24
15
15
36
21
0.6316
63.16
2250 ppm
3.35
24
6
60
6
0.9091
90.91
2750 ppm
3.44
30
0
90
0
1
100
Effect of Pare Leaf fraction n-butanol the death of Culex pipiens in shows that
increasing the concentration of n-butanol fraction is directly proportional to the number of test
animals that die and instead is inversely proportional to the number of test animals were
alive. Increasing concentrations of n-butanol fraction leaves of Pare Leaf fraction ethyl
acetate improve the ability to kill mosquito larvae of Culex pipiens. Analysis using the
method of Reed and Muench LC 99 values obtained ethyl acetate
extract fraction of
2665.63 ppm.
Based on these descriptions, it is known that the crude extract of ethanol and the
three extract fractions tested had as larvicidal activity, it is in accordance with Dalimartha
(2008), which explains that bitter melon plants can be used to kill insects. Pare leaf extract
and the fractions have the activity level is different, the difference in larvicidal activity of
extracts and fractions caused some fractionation process wherein the compound contained in
the extract chemical separation based on the degree of polarity so as to provide a
different larvicidal activity.
The content of compounds in bitter melon leaves are flavonoids, alkaloids, saponins
and terpenoids (Syamsuhidayat and Hutapea, 1991). Based on the study h acyl phytochemical
test extracts or extract fractions pare leaf by Hernawati (2006) in his research on the potential
p acre fruit (Momordica charantia L.) as a herbal ntifertilitas show that ethanol crude extract
contains alkaloids, saponins, flavonoids, and triterpenes oid. Fraction n-hexane containing
triterpenoids,flavonoids containing ethyl acetate and n -butanol containing flavonoids.
J. Islamic Pharm., an open access journal
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J. Islamic Pharm. 2017. 2(2). 13-24
21
Fortunately (1993) and Kristanti (2008) explains that the insecticide can get into the
insect's body through various ways, among others: as a stomach poison(stomatch poison) that
goes into the insect's body through the digestive tract of insects, gastrointestinal tract
mosquitoes have a structure such as a wall body . Thus the absorption of insecticides in the
digestion equals absorption of the body wall. Contact poison (contact Poisoining) that enter
through the skin or body wall, dinding body part that can absorb large amounts of
insecticide. The body wall has a base layer that are semipermeable membrane so that it can
choose the types of compounds that can pass through and the last one fumigant or breathing
into the insect's body through the respiratory system. The respiratory tract is called the
trachea. Air and oxygen enter the trachea by diffusion assisted by abdominal
movement. Oxygen will be directly related to the network. Insecticides enter the respiratory
system in the form of gas or fine grains that are brought to the living tissue.
Function alkaloids, triterpenoids, saponins and flavonoids in the leaves of bitter
melon can hinder eating larvae (antifedant). These compounds can enter the digestive through
immersion extract concentration ingested by the larvae of the mosquito Culex pipiens, then
enter the digestive tract and are absorbed by the intestinal wall and then circulate along the
blood that would interfere with the metabolism of the body of the mosquito larvae Culex
pipiens that lack of energy for activities that lead to Mosquitoes finally die. The workings of
these compounds is to act as stomatch poisoning or stomach poison. Therefore, if these
compounds are included in the larval body, ala t be disturbed digestion (Cahyadi, 2009).
CONCLUSION
The crude extract of bitter melon leaves ethanol and fractions have an activity as larvicides
against the mosquito larvae Culex pipiens the LC 99 value of each is at 7416.52 ppm ethanol
extract, extract n -heksana fraction 2564.48 ppm, extract ethyl acetate fraction 2702.09 ppm
and extract n -butanol fraction 2665.63 ppm.
ACKNOWLEDGEMENTS
The development of this plant needs to be done because this plant is easy to find and
the number is many. E kstrak and pare leaf extract fraction having as larvicidal
activity. Extract fraction n -heksana known to have the most excellent larvicidal activity, these
results can be used as a reference in use are the most excellent naan extract as larvicides for
further research so that the development of further research on bitter melon leaves that leads
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J. Islamic Pharm. 2017. 2(2). 13-24
22
to the use of these crops is necessary, the isolation of pure compound From leaf pare is a way
of development further to do in the future so that it can be used as a natural larvicidal derived
from leaf extracts of bitter melon (Momordica charantia L.).
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J. Islamic Pharm., an open access journal
ISSN : 2527-6123